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Identification of genes associated with mutacin I production in Streptococcus mutans using random insertional mutagenesis

机译:使用随机插入诱变,在链球菌Mutans中产生与Mutacin I的基因鉴定

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Streptococcus mutans is a major pathogen implicated in dental caries. Its virulence is enhanced by its ability to produce bacteriocins, called mutacins, which inhibit the growth of other Gram-positive bacteria. The goal of this study is to use a random insertional mutagenesis approach to search for genes that are associated with mutacin I production in the virulent strain UA140. A random insertional mutagenesis library consisting of 11?000 clones was constructed and screened for a mutacin-defective phenotype. Mutacin-defective clones were isolated, and their insertion sites were determined by PCR amplification or plasmid rescue followed by sequencing. A total of twenty-five unique genes were identified. These genes can be categorized into the following functional classes: two-component sensory systems, stress responses, energy metabolism and central cellular processes. Several conserved hypothetical proteins with unknown functions were also identified. These results suggest that mutacin I production is stringently controlled by diverse and complex regulatory pathways.
机译:链球菌变异是一种主要的病原体,涉及龋齿。其毒力通过其产生含有细菌素的能力,称为mutacin粉丝,其抑制其他革兰氏阳性细菌的生长。本研究的目的是使用随机插入诱变方法来搜索与毒性菌株UA140中的Mutacin I产生相关的基因。构建并筛选由11 000克隆组成的随机插入诱变文库,并筛选用于更新缺陷的表型。分离有缺陷的克隆,通过PCR扩增或质粒救援,然后进行测序测定它们的插入位点。鉴定了总共二十五个独特基因。这些基因可以分为以下功能等级:双组分感官系统,应力响应,能量代谢和中央蜂窝过程。还鉴定了几种具有未知功能的保守假设蛋白。这些结果表明,由多元化和复杂的调节途径严格地控制Mutacin I的生产。

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